Post-pulse addition of trans-cyclohexane-1,2-diol improves electrotransfer mediated gene expression in mammalian cells

• Trans-cyclohexane-1,2-diol (TCHD) collapses the permeability barrier of the nuclear pore complex.
• TCHD improves expression in gene electrotransfer.
• Post pulse TCHD addition is the most effective protocol.
• TCHD does not affect the cell viability when coupled to electrotransfer.

Electric field mediated gene transfer is facing a problem in expression yield due to the poor transfer across the nuclear envelope. Trans-cyclohexane-1,2-diol (TCHD) was shown to significantly increase chemically mediated transfection by collapsing the permeability barrier of the nuclear pore complex. We indeed observed a significant increase in expression by electrotransfer when cells are treated post pulse by a low non toxic concentration of TCHD. This was obtained for different pulsing conditions, cell strains and plasmid constructs. An interesting improvement in cell viability can be obtained. This can significantly enhance the non-viral gene electrical delivery.

1-Electric field induces permeabilisation of the plasma membrane in the areas facing the electrodes. Negatively charged plasmid DNA is pushed by electrophoretic forces toward the permeabilized membrane facing the cathode.2-plasmid DNA migrates through the cytoplasm where it is exposed to nuclease.3-TCHD collapses the permeability barrier of the nuclear pore complex facilitating the entrance of the plasmid in the nucleus.Figure optionsDownload as PowerPoint slide